两种人IgM抗脂质A单克隆抗体结合特异性及功能比较。

Circulatory shock Pub Date : 1994-12-01
M A Wisniewski, M Kazemi, I S Fang, L S Knight, C C Huntenburg, J E Bubbers, M J Schneidkraut
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引用次数: 0

摘要

两种抗脂质A单克隆抗体(mAb)- HA-1A和SdJ5-1.17.15-与其在脂质A上的抗原位点的相互作用,使用点印迹法和脂质A结构类似物以及脂质A-高密度脂蛋白(HDL)复合物进行了比较。两种单抗的反应活性均受脂肪酸侧链类型和氨基葡萄糖残基II上的磷酸基团的影响;然而,SdJ5-1.17.15的相互作用似乎更明显地受到脂肪酸侧链的影响。确定了这些抗原差异的生物学意义。用SdJ5-1.17.15预孵育的大肠杆菌055:B5脂多糖(LPS)激发的人外周血单核细胞(hPBMC)释放的肿瘤坏死因子- α (tnf - α)和白细胞介素-1 β (IL-1 β)显著低于暴露于载体预孵育LPS的人外周血单核细胞(hPBMC)。HA-1A不减弱两种细胞因子的体外释放。还评估了两种单抗对LPS体内毒性的中和能力。用SdJ5-1.17.15预先孵育的大肠杆菌055:B5给药的大鼠24小时死亡率显著降低。HA-1A没有降低体内死亡率。因此,抗脂质A单抗与抗原的反应性优先受到脂质A片段上不同残基的影响。因此,SdJ5-1.17.15和HA-1A在生物活性上的差异可能部分归因于它们在脂质A上的识别位点的差异。
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Comparison of binding specificity and the function of two human IgM anti-lipid A monoclonal antibodies.

The interactions of two anti-lipid A monoclonal antibodies (mAb)--HA-1A and SdJ5-1.17.15--with their antigenic sites on lipid A, were compared using a dot-blot assay and lipid A structural analogues, as well as lipid A-high-density lipoprotein (HDL) complexes. The reactivities of both mAb were affected by the type of fatty acid side chains and by the phosphate group on the glucosamine residue II; however, the interaction of SdJ5-1.17.15 appeared to be more markedly affected by the fatty acid side chains. A determination of the biological significance of these antigenic differences was made. Human peripheral blood mononuclear cells (hPBMC) challenged with Escherichia coli 055:B5 lipopolysaccharide (LPS) pre-incubated with SdJ5-1.17.15 released significantly less tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta), compared to hPBMC exposed to vehicle preincubated LPS. HA-1A did not attenuate the in vitro release of either cytokine. The ability of both mAb to neutralize the in vivo toxicity of LPS was also evaluated. Rats administered E. coli 055:B5 pre-incubated with SdJ5-1.17.15 had a significantly reduced 24-hr mortality rate compared to vehicle controls. HA-1A did not attenuate the in vivo mortality rate. Therefore, the reactivity of anti-lipid A mAb with the antigen is preferentially affected by different residues on the lipid A moiety. Thus, the differences in biological activity seen with SdJ5-1.17.15 and HA-1A may be due in part to differences in their recognition sites on lipid A.

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Comparison of binding specificity and the function of two human IgM anti-lipid A monoclonal antibodies. Hypertonic saline/dextran versus lactated Ringer's treatment for hemorrhage in dehydrated swine. Lysozyme regulates LPS-induced interleukin-6 release in mice. Liver oxygen uptake dependence and mitochondrial function in septic rats. Efficacy of continuous arteriovenous hemofiltration in endotoxic shock.
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